Isolation and characterization of recombinant eel growth hormone expressed in Escherichia coli.
To obtain information about the microheterogeneity of recombinant protein, recombinant eel growth hormone II (EGH) analogues expressed in Escherichia coli were isolated and characterized. The modification was classified into three types: monodeamidation of Asn, oxidation of Met and N-terminal formylation. Monodeamidated EGH was isolated by ion-exchange chromatography. The major deamidation site (Asn 147) was determined by peptide mapping using the substrate specificity of trypsin. Oxidized EGH and N-terminal-formylated EGH were isolated by reversed-phase high-performance liquid chromatography. Oxidized EGH was identified by amino acid composition analysis and N-terminal-formylated peptide by mass spectrometry.
Sugimoto
S
S
Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., Japan.
Yamaguchi
K
K
Yokoo
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D004524:Eels / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D013006:Growth Hormone / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D008969:Molecular Sequence Data; D011995:Recombination, Genetic
D013006:Growth Hormone